PD Dr. Gabor MATYAS Selected scientific projects Page 1/5 SELECTED SCIENTIFIC PROJECTS (SINCE 2006) Project 2011 2014 Molecular basis of Marfan syndrome and related aortic disorders: Whole-exome sequencing and targeted therapy Marfan syndrome (MFS) is one of the most prevalent connective tissue disorders, affecting 1-3:10 000 individuals. MFS is characterized by skeletal, ocular, and cardiovascular manifestations and exhibits a broad range of severity. The phenotypic spectrum of MFS overlaps with many monogenic aortic disorders characterized by life-threatening aortic dilatation/dissection (AD), which affect(s) millions of people worldwide. Only a limited part of genes mutated in AD/MFS is known. The main aim of the proposed research project is therefore to detect novel genes and genetic modifiers implicated in the pathogenesis of AD/MFS. This aim will be achieved using whole-exome sequencing, a combination of ultrahigh-throughput next-generation sequencing and the state-of-the-art enrichment of all known human protein-coding exons and flanking canonical splice-sites. In addition to qualitative analyses, which can detect point mutations and small insertions/deletions, whole-exome sequencing data will also be evaluated/used for quantitative sequence analysis in order to detect large insertions and deletions (copy number variations). Moreover, the further aim of the proposed project is to study the effect of pharmacological substances for the therapy of true haploinsufficiency in AD, a condition in which a disease-underlying gene has only a single functional copy, instead of two copies. The outcome of this research project will provide important new insights into the molecular basis of genetically heterogeneous aortic disorders. This knowledge allows not only improved diagnostics, surgical decision, and targeted clinical management in AD/MFS patients but it will also enable genetic counseling and early (preimplantation, prenatal, and presymtomatic) molecular diagnosis in affected families. Furthermore, our research will hopefully contribute to therapeutic/pharmacological interventions for haploinsufficient AD/MFS cases and is expected to have a widespread impact by serving as a model for any other monogenic disorder with locus heterogeneity and partly unknown molecular basis. - COFRA Foundation - G & J Bangerter-Rhyner-Stiftung
PD Dr. Gabor MATYAS Selected scientific projects Page 2/5 Project 2012 2014/2015 Ehlers-Danlos Syndrom vaskulärer Typ (EDS IV) Überprüfung der Wirkung verschiedener Substanzen auf die Aorta in Mausmodellen Allgemein verständliche Zusammenfassung Das Ehlers-Danlos Syndrom vom vaskulären Typ (EDS IV) ist eine Krankheit des Bindegewebes, welches im Körper, beispielsweise in der Haut und den Wänden von inneren Organen und Blutgefässen, eine wichtige «Binde»-Funktion hat. EDS IV wird autosomal dominant vererbt, das heisst, dass Kinder von einem betroffenen Elternteil mit einem Risiko von 50 % auch an der Krankheit leiden, unabhängig vom Geschlecht. EDS IV hat eine geschätzte Häufigkeit von 1-2 Betroffenen in 100'000 Individuen, ist aber wahrscheinlich häufiger, da viele Fälle undiagnostiziert versterben. Die Symptome umfassen eine dünne, durchscheinende Haut, typische Gesichtszüge und zerreissliche Wände von Hohlorganen und grösseren Arterien, was zu gehäuften Rupturen/Rissen führt. Die schwerwiegendste Komplikation dieser Krankheit ist natürlich das Einreissen und meistens tödliche Platzen der Hauptschlagader (Aorta) und weiterer Arterien. EDS IV wird durch einen Fehler (eine Mutation) im COL3A1-Gen verursacht, welches für die Herstellung von Kollagen Typ III verantwortlich ist. Dieses Kollagen kommt vor allem in den inneren Organen vor und sorgt für deren Reissfestigkeit bzw. mechanische Stabilität. In den meisten Fällen wird im COL3A1-Gen eine Mutation gefunden, welche zu einem strukturell veränderten Kollagen Typ III Protein (Eiweiss) führt und so seine Funktion beeinträchtigt. Nur in ganz seltenen Fällen wurden Mutationen beschrieben, welche die Produktion von Kollagen Typ III «nur» mengenmässig stark reduzieren. Bis jetzt gibt es keine kausale (ursachenbekämpfende) Therapie für Patienten mit EDS IV, sondern nur Vorsichtsmassnahmen (Life Style) und Symptombehandlung. In der von uns beschriebenen Familie (Meienberg et al. 2010, Eur J Hum Genet 18:1315-1321) ist es günstig, dass es «nur» zu wenig Kollagen Typ III hat, aber keinen strukturell veränderten Anteil, der die Kollagen-Struktur stört. Dies macht eine gezielte Therapie einfacher, da «nur» die Menge von Kollagen Typ III erhöht werden muss. Für EDS IV gibt es ausserdem nur Mausmodelle, die nur zu wenig Kollagen Typ III haben. Das heisst, dass zurzeit Tierversuche zum menschlichen EDS IV nur mit diesem Gendefekt gemacht werden können. Das Hauptziel des vorliegenden Projektes ist es, anhand dieser Mausmodelle eine Therapie zu finden, die die Gesamtmenge an Kollagen Typ III erhöht und somit zu einer besseren Stabilität der Hauptschlagader führt und so das Risiko für Risse senkt. In einem zweiten Schritt wird dann geprüft, ob und wie die gefundenen Substanzen und gewonnenen Erkenntnisse auf weitere Patienten mit EDS IV und sogar auch mit verwandten Krankheiten der Hauptschlagader angewendet werden können, mit dem Ziel, möglichst vielen Menschen helfen zu können. - Wolfermann-Nägeli-Stiftung
PD Dr. Gabor MATYAS Selected scientific projects Page 3/5 Project 2008 2011 Molecular basis of Marfan syndrome: In silico, in vitro and candidate gene analyses Mutations in the FBN1 gene cause Marfan syndrome (MFS), an autosomal dominant connective tissue disorder. Recently, heterozygous mutations in the TGFBR1 and TGFBR2 genes have been reported in MFS type 2 (MFS2) and type 3 (Loeys-Dietz aortic aneurysm syndrome, LDS) as well as in familial thoracic aortic aneurysms and dissections (TAAD). The knowledge about the presence of mutations is important for presymptomatic diagnosis and genetic counseling. However, a critical issue is to identify whether a given sequence variant is a harmless polymorphism or a disease-causing mutation. For this reason, the first part of the present project focuses on the evaluation and application of computational tools to assess the pathogenic impact of FBN1 sequence variants. In the case of unclear or confusing computational predictions, in vitro tests and cosegregation analyses will also be performed. In patients with suspected MFS, mutation analyses have failed to detect FBN1, TGFBR1 or TGFBR2 involvement in at least 10%-30% of cases, suggesting that the disease-causing mutation occurs in a different gene. The aim of the second part of the present project will therefore be to identify novel genes involved in MFS by analyzing candidate genes and screening for DNA copy number variation using high-resolution SNP arrays. The results of this project will provide the basis for improved molecular diagnostics and a better understanding of the molecular pathogenesis and phenotypic variability of MFS and related disorders, which affect at least half a million people worldwide. - Jubiläumsstiftung Swiss Life - Swiss National Science Foundation
PD Dr. Gabor MATYAS Selected scientific projects Page 4/5 Project 2007 High-resolution DNA copy number analysis in Marfan syndrome-related disorders using GeneChip Mapping arrays Mutations in the FBN1 gene cause Marfan syndrome (MFS), an autosomal dominant connective tissue disorder. Recently, heterozygous mutations in the TGFBR1 and TGFBR2 genes have been reported in MFS type 2 (MFS2) and type 3 (Loeys-Dietz aortic aneurysm syndrome, LDS) as well as in familial thoracic aortic aneurysms and dissections (TAAD). Very recently, we have found that MFS-like phenotypes are caused not only by single-base substitutions (point mutations) but are also associated with large rearrangements of these genes not detectable by standard PCR-based screening assays. In addition, increasing evidences suggest that MFS-related disorders are caused by mutations in genes not yet associated with MFS-related clinical phenotypes. By using the GeneChip Human Mapping 500K arrays, the study aims (1) to narrow down the breakpoints of large deletions recently identified by Multiplex Ligation-dependent Probe Amplification (MLPA) technique, and (2) to identify genes not yet associated with MFS-related disorders by detecting DNA copy number variations in the human genome. The outcomes of the present project contribute to the better understanding of the molecular pathogenesis of MFS-related disorders and fulfil the current needs of molecular diagnostics, enabling presymptomatic diagnosis and genetic counselling in the affected families. - Swiss Heart Foundation
PD Dr. Gabor MATYAS Selected scientific projects Page 5/5 Project 2006 2007 Molecular basis of Marfan syndrome: investigation of exonic mutations affecting splicing and large deletions in the FBN1 gene Mutations in the human FBN1 gene cause Marfan syndrome (MFS), an autosomal dominant connective tissue disorder. Knowledge about FBN1 mutations is important for early diagnosis, management, and genetic counselling of MFS patients. However, a critical issue is to identify whether a given nucleotide change is a harmless polymorphism or a disease-causing mutation. While nonsense and known missense mutations suggest a causative effect, the pathogenic consequence of sporadic cases of silent and novel missense mutations can only be determined experimentally (e.g. mrna, protein, and familial cosegregation analyses). For this reason, the first part of the present project should focus on pre-mrna processing where the effect of exonic nucleotide changes can be functionally defined. Based on a large collection of exonic FBN1 mutations we will assess how and how often missense and silent mutations influence the normal splicing of the FBN1 gene. Using a transfection assay, the effect of specific splicing factors will also be investigated. Previous studies have revealed that approximately 70% of patients who fulfilled the clinical criteria of MFS have mutations in the FBN1 gene, suggesting the presence of a second Marfan locus (MFS2) and/or large deletions not detectable by currently used PCR-based assays. This leaves about 30% of MFS patients without any molecular explanation for the clinical diagnosis, prohibiting prenatal and presymptomatic diagnostics in the respective family. The aim of the second part of the present project will therefore be to develop a suitable screening method for the detection of whole-exon deletions/insertions in the FBN1 genomic DNA. Using this method, the frequency of large deletions should be assessed in DNA samples from patients in whom previous PCR-based sequencing revealed no FBN1 mutations. The outcomes of the present project would fulfil the urgent needs of molecular diagnostics and broaden our knowledge of the molecular pathogenesis of MFS, which affects at least half a million people worldwide. - Foundation for Research at the Medical Faculty of the University of Zurich - Research Funding of the University of Zurich - Swiss Heart Foundation